Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2007-10-09
2007-10-09
Horlick, Kenneth R. (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091200
Reexamination Certificate
active
10636509
ABSTRACT:
A method for orientation-directed construction of a construct including at least two nucleic acid segments of interest. Products are provided having phosphorylated blunt ends derived from the nucleic acid segments of interest. Separate ligation reactions are performed wherein in each reaction the phosphorylated blunt-ended products of two different segments obtained in the previous step are ligated to create a combined ligated sequence. PCR amplification reaction is performed using the combined ligated sequence obtained in the previous step as a template and specific enrichment primers direct the PCR amplification towards the desired orientation. Each combined ligated sequence is amplified in a separate reaction creating a combined product having phosphorylated blunt ends. The combined product is isolated and purified to have the enriched desired orientation. Self-ligation of the isolated purified combined product is performed to create a circular double-stranded DNA construct containing the desired segments properly aligned and operably linked.
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Ben-Asouli Yitzhak
Osman Farhat
Babic Christopher M
Bianco Paul D.
Fleit Martin
Fleit Kain Gibbons Gutman Bongini & Bianco P.L.
Gene Bio-Application Ltd.
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